Formation of multicellular spheroids composed of adult rat hepatocytes in dishes with positively charged surfaces and under other nonadherent environments.
Adult rat hepatocytes formed floating multicellular spheroids in primary culture in an uncoated plastic dish with a positively charged surface. Cells in the spheroids formed in such a simple way were similar to those formed in dishes coated with proteoglycan fraction isolated from rat liver reticulin fibers; in both cases, cells maintained high ability to produce albumin and poor ability to proliferate in response to epidermal growth factor. Coating dishes with albumin was also helpful in spheroid formation; coating with 2-hydroxymethyl methacrylate resulted in formation of incomplete spheroids. Elimination of serum factors was essential for the formation of spheroids; when cells were washed with serum-containing medium before seeding or if the medium was replaced with a serum-containing medium, spheroid formation was completely inhibited. Collagens, fibronectin, and laminin, all of which promote the adhesion and spreading of hepatocytes on substrates, inhibited spheroid formation. Furthermore, collagens disintegrated spheroids, and cells in the monolayer initiated proliferation. Thus, two distinct, mutually exclusive features of primary culture of adult hepatocytes apparently exist; monolayer culture with proliferative activity in an adherent environment and spheroid culture with poor proliferative activity and high albumin-producing ability in a nonadherent environment.
Koide
N
N
First Department of Internal Medicine, Okayama University Medical School, Japan.
Sakaguchi
K
K
Koide
Y
Y
Asano
K
K
Kawaguchi
M
M
Matsushima
H
H
Takenami
T
T
Shinji
T
T
Mori
M
M
Tsuji
T
T
D000418:Albumins / Q000378:metabolism; D000818:Animals; D001769:Blood; D002448:Cell Adhesion; D002450:Cell Communication; D002455:Cell Division; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D004563:Electrochemistry; D005109:Extracellular Matrix / Q000502:physiology; D005353:Fibronectins / Q000494:pharmacology; D007797:Laminin / Q000494:pharmacology; D008099:Liver / Q000166:cytology*; D008854:Microscopy, Electron; D051381:Rats; D011919:Rats, Inbred Strains; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D000418:Albumins; D005353:Fibronectins; D007797:Laminin; D012710:Serum Albumin, Bovine; D003094:Collagen